title: Proteomics approach to study the functions of Drosophila myosin VI through identification of multiple cargo-binding proteins.
abstract: Myosin VI is a molecular motor implicated in many processes, and it likely associates with a variety of cargoes that specify its functions. Although it is critical to Drosophila development, little is known about its cellular roles. To reveal its involvement in specific pathways, we sought to identify the binding partners of Drosophila myosin VI. We used affinity chromatography and mass spectrometry to discover interacting proteins, which we tested for direct binding. Using this approach, we found that the microtubule-associated protein Cornetto bound myosin VI, and we demonstrated a role for both in secretion of the lipidated morphogen Hedgehog. We also identified a number of other binding proteins, and further characterization of their interactions with myosin VI will advance our understanding of the roles of these complexes in cellular and developmental processes. Thus, our method has provided us the means to gain valuable insight into the multifaceted roles of a motor protein in vivo.
authors.affiliation: Department of Biochemistry, Stanford University, 279 Campus Drive, Stanford, CA 94305, USA.
authors.forename: Dina
authors.initials: D
authors.lastname: Finan
authors.forename: M Amanda
authors.initials: MA
authors.lastname: Hartman
authors.forename: James A
authors.initials: JA
authors.lastname: Spudich
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004330:Drosophila / Q000235:genetics / Q000502:physiology*; D029721:Drosophila Proteins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D053823:Hedgehog Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D040901:Proteomics
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D029721:Drosophila Proteins; D053823:Hedgehog Proteins; D008869:Microtubule-Associated Proteins; C448520:corn protein, Drosophila; C089785:myosin VI; C077245:hh protein, Drosophila; D018995:Myosin Heavy Chains